Aros-Calt Sandrine, Castelli Florence A, Lamourette Patricia, Gervasi Gaspard, Junot Christophe, Muller Bruno H, Fenaille François
Service de Pharmacologie et d'Immunoanalyse, Laboratoire d'Etude du Métabolisme des Médicaments, CEA, INRA, Université Paris Saclay, MetaboHUB, Gif-sur-Yvette, France.
bioMérieux S.A., Marcy l'Etoile, France.
Methods Mol Biol. 2019;1871:279-293. doi: 10.1007/978-1-4939-8814-3_18.
Staphylococcus aureus is a major human pathogen that can readily acquire antibiotic resistance. For instance, methicillin-resistant S. aureus represents a major cause of hospital- and community-acquired bacterial infections. In this chapter, we first provide a detailed protocol for obtaining unbiased and reproducible S. aureus metabolic profiles. The resulting intracellular metabolome is then analyzed in an untargeted manner by using both hydrophilic interaction liquid chromatography and pentafluorophenyl-propyl columns coupled to high-resolution mass spectrometry. Such analyses are done in conjunction with our in-house spectral database to identify with high confidence as many meaningful S. aureus metabolites as possible. Under these conditions, we can routinely monitor more than 200 annotated S. aureus metabolites. We also indicate how this protocol can be used to investigate the metabolic differences between methicillin-resistant and susceptible strains.
金黄色葡萄球菌是一种主要的人类病原体，能够轻易获得抗生素耐药性。例如，耐甲氧西林金黄色葡萄球菌是医院获得性和社区获得性细菌感染的主要原因。在本章中，我们首先提供了一个详细的方案，用于获得无偏差且可重复的金黄色葡萄球菌代谢谱。然后，通过使用亲水相互作用液相色谱法和与高分辨率质谱联用的五氟苯基丙基柱，以非靶向方式分析所得的细胞内代谢组。这些分析结合我们内部的光谱数据库进行，以尽可能高置信度鉴定出许多有意义的金黄色葡萄球菌代谢物。在这些条件下，我们可以常规监测200多种注释的金黄色葡萄球菌代谢物。我们还指出了该方案如何用于研究耐甲氧西林菌株和敏感菌株之间的代谢差异。
